Molecular diagnosis is used to detect the basic causes of disease, such as DNA or RNA molecules, in various diagnostic fields, including infectious disease diagnosis, cancer diagnosis, genetic disease diagnosis and personalized diagnosis fields. Typical molecular diagnosis techniques are PCR techniques for amplifying DNA within a short time (Saiki, R., et. al. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239, 487-91. 1998). However, in general PCR techniques, electrophoresis should be used to confirm amplified DNA. Before such electrophoresis, complicated procedures should be performed, including making agarose gel and staining DNA with EtBr or the like. For this reason, electrophoresis is not suitable for use in clinical laboratories.
Real-time PCR techniques that have recently been used are based on fluorescence, and thus electrophoresis is not required, but have a defect in that expensive instruments and expensive fluorescent reagents are used (Higuchi, R., et. al., Kinetic PCR Analysis: Real-time Monitoring of DNA Amplification Reactions. Nature Biotechnology 11, 1026-1030, 1993). In addition, these techniques have a shortcoming in that, because fluorescence wavelengths to be used are limited, it is difficult to practically perform multiplex PCR for five or more samples. For these reasons, there are many difficulties in performing inexpensive clinical tests by use of PCR techniques in laboratories, and thus PCR techniques are used mainly in large-sized hospitals such as university hospitals.
In recent years, GeneXpert system and reagents (Cepheid) for use in on-site diagnosis have been developed and marketed. However, the system and reagents are highly expensive, and thus are hardly used in general clinical tests (Helb, D., et. al., Rapid Detection of Mycobacterium tuberculosis and Rifampin Resistance by Use of On-Demand, Near-Patient Technology. J. Clin. Microbiol. 48, 229-237, 2010).
Other techniques include a nucleic acid lateral flow assay that is performed using a membrane instead of gel electrophoresis after PCR (Nucleic Acid Lateral Flow Assay)  (Aveyard, J., et. al., One step visual detection of PCR products with gold nanoparticles and a nucleic acid lateral flow (NALF) device. Chem. Commun., 41, 4251-4253, 2007). However, this nucleic acid lateral flow assay is more complicated than gel electrophoresis techniques, and thus is impossible to use in laboratories. In addition, as there are technical limitations in a view that the sequence of a probe attached to the membrane should be used so that it can bind specifically to a PCR amplification product, the universal use of the nucleic acid lateral flow assay is limited.
Under this technical background, the present inventors have found that, when a nucleic acid oligomer in an artificially synthesized PCR primer is allowed to react with a probe complementary to the nucleic acid oligomer on a membrane regardless of types on amplification product, various PCR products can be universally identified using a single type of membrane, thereby completing the present disclosure.
The information disclosed in the Background Art section is only for the enhancement of understanding of the background of the present disclosure, and therefore may not contain information that forms a prior art that would already be known to a person of ordinary skill in the art.